Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton
Dendritic spines (DS) are tiny protrusions implicated in excitatory postsynaptic responses in the CNS. To achieve their function, DS concentrate a high density of ion channels and dynamic actin networks in a tiny specialized compartment. However, to date there is no direct information on DS ionic co...
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| Format: | Article |
| Language: | English |
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Frontiers Media S.A.
2022-08-01
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| Series: | Frontiers in Molecular Neuroscience |
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| Online Access: | https://www.frontiersin.org/articles/10.3389/fnmol.2022.769725/full |
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| author | Avner Priel Xiao-Qing Dai Xing-Zhen Chen Noelia Scarinci María del Rocío Cantero Horacio F. Cantiello |
| author_facet | Avner Priel Xiao-Qing Dai Xing-Zhen Chen Noelia Scarinci María del Rocío Cantero Horacio F. Cantiello |
| author_sort | Avner Priel |
| collection | DOAJ |
| description | Dendritic spines (DS) are tiny protrusions implicated in excitatory postsynaptic responses in the CNS. To achieve their function, DS concentrate a high density of ion channels and dynamic actin networks in a tiny specialized compartment. However, to date there is no direct information on DS ionic conductances. Here, we used several experimental techniques to obtain direct electrical information from DS of the adult mouse hippocampus. First, we optimized a method to isolate DS from the dissected hippocampus. Second, we used the lipid bilayer membrane (BLM) reconstitution and patch clamping techniques and obtained heretofore unavailable electrical phenotypes on ion channels present in the DS membrane. Third, we also patch clamped DS directly in cultured adult mouse hippocampal neurons, to validate the electrical information observed with the isolated preparation. Electron microscopy and immunochemistry of PDS-95 and NMDA receptors and intrinsic actin networks confirmed the enrichment of the isolated DS preparation, showing open and closed DS, and multi-headed DS. The preparation was used to identify single channel activities and “whole-DS” electrical conductance. We identified NMDA and Ca2+-dependent intrinsic electrical activity in isolated DS and in situ DS of cultured adult mouse hippocampal neurons. In situ recordings in the presence of local NMDA, showed that individual DS intrinsic electrical activity often back-propagated to the dendrite from which it sprouted. The DS electrical oscillations were modulated by changes in actin cytoskeleton dynamics by addition of the F-actin disrupter agent, cytochalasin D, and exogenous actin-binding proteins. The data indicate that DS are elaborate excitable electrical devices, whose activity is a functional interplay between ion channels and the underlying actin networks. The data argue in favor of the active contribution of individual DS to the electrical activity of neurons at the level of both the membrane conductance and cytoskeletal signaling. |
| first_indexed | 2024-04-11T21:17:56Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 1662-5099 |
| language | English |
| last_indexed | 2024-04-11T21:17:56Z |
| publishDate | 2022-08-01 |
| publisher | Frontiers Media S.A. |
| record_format | Article |
| series | Frontiers in Molecular Neuroscience |
| spelling | doaj.art-c4d83f4bb4204abe8ff7df44a49b8c4e2022-12-22T04:02:45ZengFrontiers Media S.A.Frontiers in Molecular Neuroscience1662-50992022-08-011510.3389/fnmol.2022.769725769725Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeletonAvner Priel0Xiao-Qing Dai1Xing-Zhen Chen2Noelia Scarinci3María del Rocío Cantero4Horacio F. Cantiello5The Mina & Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, IsraelDepartment of Pharmacology, Alberta Diabetes Institute, University of Alberta, Edmonton, AB, CanadaDepartment of Physiology, University of Alberta, Edmonton, AB, CanadaLaboratorio de Canales Iónicos, Instituto Multidisciplinario de Salud, Tecnología y Desarrollo, Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET) - Universidad Nacional de Santiago del Estero (UNSE), Santiago del Estero, ArgentinaLaboratorio de Canales Iónicos, Instituto Multidisciplinario de Salud, Tecnología y Desarrollo, Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET) - Universidad Nacional de Santiago del Estero (UNSE), Santiago del Estero, ArgentinaLaboratorio de Canales Iónicos, Instituto Multidisciplinario de Salud, Tecnología y Desarrollo, Consejo Nacional de Investigaciones Científicas y Técnicas de Argentina (CONICET) - Universidad Nacional de Santiago del Estero (UNSE), Santiago del Estero, ArgentinaDendritic spines (DS) are tiny protrusions implicated in excitatory postsynaptic responses in the CNS. To achieve their function, DS concentrate a high density of ion channels and dynamic actin networks in a tiny specialized compartment. However, to date there is no direct information on DS ionic conductances. Here, we used several experimental techniques to obtain direct electrical information from DS of the adult mouse hippocampus. First, we optimized a method to isolate DS from the dissected hippocampus. Second, we used the lipid bilayer membrane (BLM) reconstitution and patch clamping techniques and obtained heretofore unavailable electrical phenotypes on ion channels present in the DS membrane. Third, we also patch clamped DS directly in cultured adult mouse hippocampal neurons, to validate the electrical information observed with the isolated preparation. Electron microscopy and immunochemistry of PDS-95 and NMDA receptors and intrinsic actin networks confirmed the enrichment of the isolated DS preparation, showing open and closed DS, and multi-headed DS. The preparation was used to identify single channel activities and “whole-DS” electrical conductance. We identified NMDA and Ca2+-dependent intrinsic electrical activity in isolated DS and in situ DS of cultured adult mouse hippocampal neurons. In situ recordings in the presence of local NMDA, showed that individual DS intrinsic electrical activity often back-propagated to the dendrite from which it sprouted. The DS electrical oscillations were modulated by changes in actin cytoskeleton dynamics by addition of the F-actin disrupter agent, cytochalasin D, and exogenous actin-binding proteins. The data indicate that DS are elaborate excitable electrical devices, whose activity is a functional interplay between ion channels and the underlying actin networks. The data argue in favor of the active contribution of individual DS to the electrical activity of neurons at the level of both the membrane conductance and cytoskeletal signaling.https://www.frontiersin.org/articles/10.3389/fnmol.2022.769725/fullhippocampushippocampal neuronsdendritic spinesNMDA receptorsynapseelectrical oscillations |
| spellingShingle | Avner Priel Xiao-Qing Dai Xing-Zhen Chen Noelia Scarinci María del Rocío Cantero Horacio F. Cantiello Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton Frontiers in Molecular Neuroscience hippocampus hippocampal neurons dendritic spines NMDA receptor synapse electrical oscillations |
| title | Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton |
| title_full | Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton |
| title_fullStr | Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton |
| title_full_unstemmed | Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton |
| title_short | Electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton |
| title_sort | electrical recordings from dendritic spines of adult mouse hippocampus and effect of the actin cytoskeleton |
| topic | hippocampus hippocampal neurons dendritic spines NMDA receptor synapse electrical oscillations |
| url | https://www.frontiersin.org/articles/10.3389/fnmol.2022.769725/full |
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